1. Field of the Invention
The field of this transformation.
2. Background of the Invention
The introduction of an exogenous nucleic acid sequence (e.g. DNA) into a cell, a process known as xe2x80x9ctransformation,xe2x80x9d plays a major role in a variety of biotechnology and related applications, including research, synthetic and therapeutic applications. Research applications in which transformation plays a critical role include the production of transgenic cells and animals. Synthetic applications in which transformation plays a critical role include the production of peptides and proteins. Therapeutic applications in which transformation plays a key role include gene therapy applications. Because of the prevalent role transformation plays in the above and other applications, a variety of different transformation protocols have been developed.
In many transformation applications, it is desirable to introduce the exogenous DNA in a manner such that it is incorporated into a target cell""s genome. One means of providing for genome integration is to employ a vector that is capable of homologous recombination. Techniques that rely on homologous recombination can be disadvantageous in that the necessary homologies may not always exist; the recombination events may be slow, etc. As such, homologous recombination based protocols are not entirely satisfactory.
Accordingly, alternative viral based transformation protocols have been developed, in which a viral vector is employed to introduce exogenous DNA into a cell and then subsequently integrate the introduced DNA into the target cell""s genome. Viral based vectors finding use include retroviral vectors, e.g. Moloney murine leukemia viral based vectors. Other viral based vectors that find use include adenovirus derived vectors, HSV derived vectors, sindbis derived vectors, etc. While viral vectors provide for a number of advantages, their use is not optimal in many situations. Disadvantages associated with viral based vectors include immunogenicity, viral based complications, and the like.
Accordingly, there is continued interest in the development of additional methods of integrating exogenous nucleic acid into the genome of a target cell for use in transformation protocols. Of particular interest is the development of a non-viral in vivo nucleic acid transfer protocol that provides for stable genome integration by a mechanism other than homologous recombination.
3. Relevant Literature
Patent applications of interest include: WO 98/40510 and WO 99/25817. Also of interest are: Dawson and Finnegan, Nat. Biotechnol. (1998) 16:20-21; Ivics et al., Cell (1997) 91: 501-510; Ivics et al., Proc. Nat""l Acad. Sci. USA (1996) 93:5008-5013; Luo et al., Proc. Nat""l Acad. Sci USA (1998) 95:10769-10773; Schouten et al., Nuc. Acids Res. (1998) 26:3013-3017; Zhang et al., Nuc. Acids Res. (1998) 26:3687-3693.
Methods and compositions for introducing an exogenous nucleic acid into the genome of at least one cell of a multicellular organism are provided. In the subject methods, a Sleeping Beauty transposon that includes the exogenous nucleic acid is administered to the multicellular organism along with a source of a Sleeping Beauty transposase activity. Following administration, the exogenous nucleic acid is integrated into the genome of at least one cell of the multicellular organism. The subject methods find use in a variety of different applications, including the in vivo transfer of a gene into a target cell e.g. for use in gene therapy applications.